Fractionation of the naringinase complex from Aspergillus terreus by dye affinity chromatography

Affinity chromatography with immobilised triazine dyes was used to separate the main enzymes present in the naringinase complex produced by Aspergillus terreus CECT 2663. One alpha-L-rhamnosidase and two beta-D-glucosidases (betaG1 and betaG2) were separated by a simple two-step procedure involving chromatography with Red HF-3B immobilised on Sepharose 4B first at pH 5.5 and then at pH 4.7. Maximum activity of the beta-D-glucosidases was from pH 4 to 6 and at 65 degreesC. Both glucosidases were active on p-nitrophenol glucoside and prunin with respective Km values of 1.9 mM and 1.6 mM for betaG1 and 2.1 mM and 0.25 mM for betaG2. Only betaG1 hydrolysed cellobiose (Km = 5.7 mM).